With evolution and advances in breeding techniques of grapes

1. Course : FSC 601
ADVANCES IN BREEDING OF GRAPES
by
SHRUTHI.S2020632002
I Ph.D
Department of Fruit Science.

2. » Origin - Asia Minor (Black to Caspian sea)
» Cultivation/Study of Vine - Viticulture
» Inflorescence – Panicle
» Fruit – Berry
» Edible portion – Pericarp & Placenta
» Science of wine making – Oenology
» Introduced by – Christian missionaries in 1832 from France
» Grape aroma & flavour – Methyl anthranilate
» Resveratrol – Anti oxidant ( red wine )
» Predominant sugar – Fructose
» Red colour of grape - Anthocyanins
» Flowers – Laterally on axial of leaves on current season growth
» Growth curve - Double sigmoid growth curve
» Maharashtra - Max area and production

4. EVOLUTIONARY MECHANISM:
Local Varieties
domestication 4000 B.C. geographical spread USA : 17th Century
experimental crossing
Sterile Hybrids 2n = 39 13Ⅱ: 13 Ι
Repeated Crossing (America and Asia)
Natural and Experiment
Hybrid Varieties vinifera backcross
widely adapted
auto - 4x allo – 4x
little use potential
VITIS
60 ecospecies
Dioecious, interfertile
2n = 2x = 38
North and Central America, Asia
Wild vinifera : Central Asia
MUSCADINIA
3 spp
2n = 2x = 40
North America
Wild rotundifolia
Special varieties
often hermaphrodite
own roots, later as scions.
Wine Table and Dried
Acid sweet
Seedy less seedy
Juicy firm flesh
parthenocarpy

5. The distribution and spread of vines through the ages (from Pretorius, 2000).

7. » Stage 1 is “management,” which reflects human stewardship and
harvesting of wild plant populations, presumably by hunter-
gatherers, prior to their purposeful cultivation. The recognition of
this stage is relatively nascent and somewhat speculative, but it
conforms to a growing body of evidence that humans greatly
affected flora, as well as fauna, prior to the onset of agriculture.
» Stage 2 is purposeful cultivation, a process that undoubtedly
included selection for desirable traits, like fruit size and taste, but
also unintended selection for traits associated with growth
conditions and responses to stresses. Because it is likely that only
a subset of wild populations was cultivated, this stage often
features a dramatic domestication bottleneck that reduces and
repatterns genetic diversity relative to the wild ancestor.
» Stage 3 is geographic expansion, which occurs when the incipient
crop is dispersed to new locations. These new locations present
novel biotic and abiotic stresses that further drive crop adaptation
Interestingly, some of this adaption may be due to hybridization
with—and introgression from—locally adapted wild plants.
» Stage 4 is modern, deliberate breeding, a process that has
occurred only over the last few hundred years for most crops
(Meyer and Purugganan 2013)
sylvestris

8. л One of the major phenotypic shifts that occurred during grapevine domestication was a transition in the
mating system. At some point during the domestication process, grapes transitioned from dioecy (i.e.,
separate male and female individuals) in sylvestris to hermaphroditic individuals in sativa. This shift is
particularly dramatic given that all extant wild Vitis species are dioecious. Hence, dioecy has been
maintained since the origin of the genus, which is estimated to have occurred from * 18Mya (Wan et al.
2013) to * 39Mya (Liu et al. 2016).
л A switch to hermaphroditism provides immediate advantages for cultivation. In hermaphrodites, all
individuals can contribute to fruiting and to pollination. In contrast, only half of the population bears fruit in
a dioecious species.
л In agricultural settings, dioecy means that most males must be removed from the fields as soon as they
can be identified. For that reason, dioecy is particularly disadvantageous for agricultural productivity in
perennial crops (other e.g., date palms, persimmons, and kiwifruit), where first flowering takes many years
and sex can be identified only after first flowering. To circumvent this problem, substantial efforts have
been focused on identifying molecular markers that allow for earlier gender identification.
л The timing of the switch to hermaphroditism in grapes is unknown, but we assume it occurred early and
rapidly during stage 2 of domestication.
FEMALE
HERMAPHRODITE
MALE

9. Based on the fertility
level of pollen, grape
can be grouped in to 3
categories.
Self fertility group have
erect stamens with
oblong blunt and quite
symmetrical pollen, size
of pollen grain is very
small i.e., 0.0254mm in
diameter.
However, self-sterile
and partly self-sterile is
characterized by the
presence of impotent
pollen.
According to Jauhari et
al.(1969) peak time of
anthesis under North
Indian conditions was
between 7.00 to 9.00am.
Self pollination is rule in
V. vinifera grape, but
cross pollination is
required in reflexed
anther cultivars.

10. DOMESTICATED
WILD

11. Vitis vinifera (Common Grape)
CHARACTERISTICS : Flaky bark. leaves are alternate,
palmately lobed long and broad. The fruit is a berry can
be green, red, or purple (black)
Vitis labrusca (Fox grape)
CHARACTERISTICS : high in phenolic compounds,
Vitis aestivalis (Summer grape)
CHARACTERISTICS : vigorous vine, the leaves long,
sub orbicular, fruit is a small grape .
IMPORTANT SPECIES :

12. Vitis rupestris (Sand grape)
CHARACTERISTICS : used in breeding and as
a root stock
Vitis cadicans (Mustang grape)
CHARACTERISTICS : dark purple berries thick
outer layer of flesh and on average contain four
seeds
Vitis riparia (Frost grape)
CHARACTERISTICS : resistance to
phylloxera and adaptation to variant soil
Types

14. GENETIC RESOURCES:
Genetic resources can be divided into 4 types;
» Wild species, (The wild species are potential sources for various traits, the
Vitis genes has approximately 60 inter fertile species. North-western
Himalaya is also inhabited with two edible wild species viz. V. parviflora and
V. lanata and numerous natural hybrids of the two species which are locally
grown.)
» Old traditional cultivars,
» New cultivars and
» Breeding lines
~More than 10,000 named varieties of grapes in different countries
~Chadha and Randhawa described 130 grape varieties available in India.
~In India, germplasm are being maintained in field gene banks.
~Number of existing germplasm at IIHR, Bangalore is 616 (Anon., 1996).
~At NRCG Pune 524 accessions.

16. Network of first-degree relationships among common grape cultivars.
Sean Myles et al. PNAS 2011;108:9:3530-3535
©2011 by National Academy of Sciences

18. • Cytogenetics is the branch of genetics that studies the structure of DNA
within the cell nucleus. This DNA is condensed during cell division and form
chromosomes. The cytogenetics studies the number and morphology of
chromosomes. Using chromosome banding techniques (classical
cytogenetics) or hybridization fluorescently labeled probes (molecular
cytogenetics).
• The first cytogenetic study of these hybrids was by Patel and Olmo (1955)
who found F1 hybrids had a chromosome number of 2n = 39 and that
functional gametes were n=20 ± 1, with 20 the most frequent.
• When studying meiotic chromosome pairing of 3 hybrids they found 13
bivalents (range 6-18); 10 univalents (3-15); and 2 quadrivalents (0-4),
leading them to propose a genomic constitution of 13RrRr +7AA for V.
rotundifolia, 13RVRV +6BB for V. vinifera, and 13RrRv +6A + 7B for the F1
hybrid.
• The R chromosomes are homologous between V. vinifera and
V.rotundifolia and the A and B represent two genomes from different,
unknown ancestral species. They also state sterility is not only due to
chromosome differences but mainly to abnormal pairing and irregular
distribution.
• Meiotic irregularities, such as lagging chromosomes, bridge-fragment
formation, and chromosome elimination were common in hybrids,
indicating presence of structural dissimilarities like inversions and
translocations in the common genome.
CYTOGENETICS:

19. Crossability of V. vinifera and V. rotundifolia and their
hybrids.
_______ Successful fruit set,
+ + + failure of fruit set,
- - - some fruit set, but seeds in viable,
+ indicates maternal parent.
Cross-hatched circles - V. rotundifolia cytoplasm, clear
circles - V. vinifera cytoplasm.
Three essential facts are established concerning
crossability of the two species:
1. Whenever the maternal diploid plant contains two
chromosomal complements of V. rotundifolia, the
generative nucleus of V. vinifera or VR hybrids fail
to fertilize the V. rotundifolia egg. If the maternal
diploid contains only one or a partial chromosomal
complement of V. rotundifolia, fertilization succeeds
with V. vinifera and VR pollen.
2. In allotetraploids at the ratio 2 : 2 of V. vinifera-
rotundifolia complements in the maternal plant,
pollen of autotetraploid V. vinifera and VVRR
hybrids fertilize the egg. With 4 V. rotundifolia
complements in the female no setting was obtained
with tetraploid V. vinifera or VVRR hybrids.
3. In a maternal plant with 2 : 2 VR complements,
pollen of diploid V. vinifera fertilizes the egg. Pollen
of VVR fertilizes diploid but not tetraploid V.
rotundifolia. The cross RR X VVRR produced berry
set but all seeds were found to be floaters and
nonviable.

20. CHEMOTAXONOMY:
These are typical phytochemicals found in grape:
phenolic acids, flavanols, flavon-3-ols, myricetin,
peonidin, flavonoids, resveratrol, quercetin,
tannins, anthocyanins, kaempferol, cyanidin, ell
agic acid, proanthocyanidins.

21. INHERITANCE PATTERN:
In grapevines, inheritance and genetic analyses have been carried out only for a limited number
of traits because it has a long life cycle, a large number of chromosomes, partial sterility of ovules, and
low seed germination.
Downy mildew resistance
(1) a single gene for the hypersensitive reaction of stomatal tissues, for which resistant species are
homozygous dominant, and the susceptible V. vinifera is homozygous recessive,
(2) polygenes for inhibition of the mycelium growth in plant tissues. Broad sense heritability for downy
mildew resistance was estimated at 0.83 to 0.94, suggesting minimum environmental effects, while
narrow sense heritability was 0.26 to 0.39.
Powdery mildew resistance
The narrow sense heritability estimate for this trait ranged from 0.31-0.51, suggesting quite a high
contribution of additive genetic variance.
Botrytis resistance - synthesis of phytoalexin stilbene
Resistance to anthracnose - controlled by two dominant genes for susceptibility (An1 and An2) and
a single dominant resistance gene (An3).
Resistance to phylloxera, an insect pest, is controlled by multiple genes.

22. Two types of seedlessness are found in grapevine:
(1) stenospermocarpic, the seeds abort while still small and soft, (utilized in table
grape improvement)
(2) parthenocarpic, the seeds do not develop at all.
» The seedlessness is related to three independent recessive genes regulated by a
dominant inhibitor locus, SdI (Seed Development Inhibitor).
» Two pairs of genes affect fruit color with epistatic action: B, a dominant gene for black
fruit, and R, a dominant gene for red fruit. The white-fruited grapes are recessive for
both genes.
» Similarly, the composition of fruit anthocyanins is controlled by 2 genes: G for
diglucosides or g for monoglucosides, and O for triphenols or o for diphenols.
» For fruit aroma, muscat flavor is controlled by 5 dominant complementary genes,
methyl anthranilate is controlled by 3 dominant complementary genes, while volatile
ester levels are determined by 2 genes. Several quality traits have generally high
heritabilities: cluster compactness (0.55), berry weight (0.49), skin texture (0.75) and
pulp texture (1.0)

23. The root stocks which are tolerant to any
biotic or abiotic stresses are susceptible
to some other problems .
The released rootstocks are not suitable to
all types of environmental conditions and
soils.
Pollen storage for hybridisation technique.
DIFFICULTIES
IN GRAPE
BREEDING :

24. BREEDING OBJECTIVES
To develop
varieties with
medium vigour
and productive
basal bud,
which can be
trained on
head or pandal
system of
training.
To develop high
yielding and high
quality varieties
with increased
fruitfulness of
basal buds, less
degree of apical
dominance,
suitability for
different purpose
such as table,
raisin, wine and
juice and
resistance to
diseases.
To develop
early maturing,
seedless and
sweet cultivars
for table
purpose.
To induce
resistance to
anthracnose,
Phylloxera and
chaffer beetle.
To develop
root stocks
resistant to
salinity,
nematodes
and drought.

25. BREEDING
METHODS
Introduction &
selection
Hybridisation
Clonal
selection
Mutation breeding
Polyploidy breeding

26. INTRODUCTION:
USA- Thompson
Seedless, Perlette,
Beauty Seedless,
Flame Seedless
1281, Dogridge,
Pride, Dixie,
Wedor and Black
Corianth
USSR - Kishmish
Beli and Kishmish
Chorni (Singh and
Singh,1972).
Australia - Ruby
Seedless, Gordo
Blano, Reisling,
MS 18-55,MS 19-
77, MS 16-2,
Wortly Hall
hybrids
Canada -0912
Horizon (SW) ,
0913 Leon Millet,
Foch and 0912
Swanson Red for
wine, raisin and
table purposes.

27. Perlette
Flame seedless
Tas-e-Ganesh Sharad seedless
Bangalore Blue

28. Manjari Shayama

29. Chardonay
Pinot Noir
Cabernet Franc Sauvignon Blanc
Anab-e-Shahi

30. Cheema
sahabi
S – 49 at
Punjab is
based on
this method
only .
Some promising
seedlings from open
pollinated
population of and
Pandhari Sahebi
Kabul Manuka were
also selected.
SELECTION FROM OPEN
POLLINATED SEEDS :

31. 3. CLONAL SELECTION :
INSTITUTE CLONAL SELECTION FROM
IARI Pusa seedless Thomson seedless
Haryana Agriculture
university
HS – 37 – 6
Early maturing clone
Perlette
NRS Grapes , Pune Manjari naveen Centennial seedless
NRS Grapes , Pune Krishmish Rozavis White Kishmish Rozavis
Other examples :
Tas – a – ganesh - elongated berries .
Sonaka - much elongated berries than Tas –a–ganesh.
Rao sahebi – Cheema sahebi
Dilkush – A bud spot of Anab – e – Shahi .

33. » In India, hybridization work was started in 1958 at IARI, New Delhi. The purpose of
hybridization at IARI, New Delhi was to develop early maturing, high yielding, better
quality seedless varieties with resistant to biotic stresses.
» However, IIHR, Bangalore, started breeding programme in 1968, with objective to
develop superior varieties for table, raisins, wine and juice.
» On the basis of types of parent used, it can be grouped into two.
(a) Interspecific / Intergeneric hybridization
(b) Interspecific or Intervarietal hybridization.

34. CULTIVARS PARENTS INSTITUTION CHARACTERS
Pusa Urvashi Hur x Beauty Seedless IARI, New Delhi Tolerant to anthracnose
Pusa Navarang Madeline Angevine x Ruby
Red
IARI, New Delhi Tenturier variety
Arkawati Black Champa x Thomson
Seedless
IIHR, Bangalore, 1980 TSS 22-25%, seedless berry,
suitable for raisin making,
Arka Kanchan Anab-e-Shahi x Queen of
the
Vine Yards
IIHR, Bangalore, 1980 Bunch is large, golden
yellow colour berry, TSS 17-
20˚B, having muscat flaovur,
seeded cultivar.
Arka Shyam Bangalore Blue x Black
Champa
IIHR, Bangalore TSS 20-25ºB, resistant to
anthracnose disease.
Arka Shweta or
Shweta Seedless
Anab-e-Shahi x Thompson
Seedless
IIHR, Bangalore, 1994 TSS 18-19ºB, acidity 0.5-
0.6%, berry greenish yellow.
Arka Hans Bangalore Blue x Anab-e-
Shahi
IIHR, Bangalore, 1980 TSS 18-21ºB, suitable for
making quality wine,
resistant to anthracnose

35. CULTIVARS PARENTS INSTITUTION CHARACTERS
Arka Chitra Angur Kalan x Anab-Shahi IIHR, Bangalore Table purpose, Tolerant to
powdery mildew
Arka Krishna Black Champa x
Thompson Seedless
IIHR, Bangalore Mainly for juice purpose
Arka Majestic Angur Kalan x Black
Champa
IIHR, Bangalore Tolerant to anthracnose
disease, table purpose,
head system of training
Arka Neelmani Black Champa x
Thompson
Seedless
IIHR, Bangalore Good for red wine
Arka Soma Anab –e-Shahi x Queen of
the Vine Yards
IIHR, Bangalore Suitable for making quality
wine, tolerant to
anthracnose, powdery
mildew, downy mildew
Arka Trishna Bangalore Blue x Convent
large black
IIHR, Bangalore Wine purpose

36. MUTATION BREEDING
Mutation breeding may be attempted as a complementary tool in grape
breeding for one or more important characters, without altering the whole
genetic setup. The important mutagens used in grape breeding are physical
mutagens (χ ray and γ rays ) and chemical mutagens (Ethyl Methane Sulphonate
(EMS),N-Nitroso-N-Methyl Urethane (NMUT) and N- Nitrose -N-Methyl-Urea
(NMU).
CULTIVAR FEATURES
New Perlette (Loose Perlette) comparatively loose bunch than perlette,
Self
thinning property
Red Niagara having red fruit from Niagara
Robin Cardinal an early maturing variety from Cardinal

37. Polyploidy breeding:
* Polyploidy breeding has immense importance in the improvement of table grapes.
* The chief benefit from polyploidy is the increase in berry size.
* Autotetraploid are found to be considerably sterile and are less productive than the
parents.
* The crossing of diploid with induced tetraploids may help in evolving new triploid
seedless grapes.
* The triploids are highly sterile.
* Allotetraploids even between infertile species have been more desirable as
commercial varieties. Colchicine is generally used as an aqueous solution of 0.25-
5.0% with 5-10% glycerine to induce polyploidy.
* Marvel Seedless from Delight, Early Giant from Campbell, Muscat Common Hall
from Muscat Alexandria, Black King from Campbell, Wallis Giant from Concord, Case
from Sultana …etc. are important examples of polyploidy.

38. Embryo rescue
technique:
Seedlessness is a
desirable character for
table and raisin grapes.
Inheritance of
seedlessness is
postulated to
depend on two
complementary
recessive genes
and only about
7.5% of the total
progeny from
crosses between
Seeded x
Seedless grapes
produced fruits
without noticeable
seed traces.
The embryo rescues
theoretically increases the
proportion of seedless
progeny as it makes
possible to cross two
seedless varieties.
Ovules are
excised
before
abortion and
are cultured
on either filter
paper in liquid
medium or
solid medium.
Biotechnological tools:

40. Genetic engineering / plant
transformation:
Some encouraging preliminary
results have been obtained on Agro
bacterium mediated transformation
of grape vines.
But the production of genetically
transformed grape vines which
express a marker gene is yet to be
reported.

41. • Protoplasts are of great importance as tool
for genetic amelioration and somatic
hybridization.
• But regeneration of grape vines from
protoplasts has not yet been successful
Protoplast
culture:
• Anther culture can result into haploid grape vines
which can then be developed into homozygous
diploids by doubling chromosomes.
• These homozygous diploids will be very useful for
producing F1 hybrids and for making genetic
studies.
• But there is low success rate of regeneration of
grape vines from anther and only one case of
haploid has been reported in grape.
Anther
culture:

42. ROOTSTOCK BREEDING :
Very little work has been
done in India on breeding
cultivars showing resistance
to biotic and abiotic stresses.
Species like V. berlandieri, V.
riperii, V. candicans, V. longii
and V .champini tolerate
excessive salt in the soil and
thus can be used in
improvement programmes
oriented to breed salt
resistant types (Jindal and
Srinivasa Rao, 1988).
The two rootstocks (Dog
Ridge and Salt Creek) impart
nematode and salt resistance
to the grafts. Another
rootstock Hormony (an open-
pollinated seedling of the
cross Dog Ridge x Couderc
1613) has also been found
highly effective in imparting
nematode resistance.

43. Rootstocks suggested based on the global data:
S.No Situation/problem Rootstock
1 Water shortage 1103 P, 140 RU, 110 R, 420 A, SO 4, 99 R, St.
George, Dogridge
2 Soil EC more than 2 m mohs/cm and water EC
more than 1 m mohs/cm (Slightly Saline Soil)
Ramsey, Dogridge, 140 RU, 99 R, 110 R.
3 Soil ESP more than 15 per cent and/or water SAR
more than 8.
(Slightly Alkaline Soil)
140 RU, 1613, Ramsey, Dogridge.
4 Free calcium content of soil is more than 12% 140 RU, SO 4, 420 A.
5 Chloride content of water is more than 4
meq/litre
Ramsey, Dogridge B, 140 RU. Teleki 5-C
6 Poor vigor of the variety without any soil/water
problem
Dogridge, St. George, SO 4, 140 RU.
7 For increased nitrogen, potassium uptake. Dogridge, St. George, 34 EM, Ramsey.
8 For increased bud break 1613, B2-56

44. REFERENCES:
1. Alleweldt G, Possingham JV. Progress in grapevine breeding. Theor Appl
Genet. 1988;75:669–673. doi: 10.1007/BF00265585. [CrossRef] [Google Scholar].
2. O.I.V. Descriptor list of grape vine varieties and Vitis species. Paris: Office International de
la Vigne et du Vin. Code number 225 (1983)
3. Breeding of Fruit Crops – Dr. P.K. Ray.
4. Tropical and Subtropical Fruit Crops – T.K. Bose.
5. NRCG, Pune (Vision for Improvement – 2015 - 2020)

45. THANK YOU..